Knowledge of the biochemical pathways by which cells detect and respond to stimuli is important for the discovery, development, and correct application of pharmaceutical products. Cellular physiology involves multiple pathways, which have complex relationships. For example, pathways split and join; there are redundancies in performing specific actions; and response to a change in one pathway can modify the activity of another pathway, both within and between cells. In order to understand how a candidate agent is acting and whether it will have the desired effect, the end result, and effect on pathways of interest is as important as knowing the target protein.
BioMAP® methods of analysis for determining the pathways affected by an agent or genotype modification in a cell, and for identifying common modes of operation between agents and genotype modifications, are described in International Patent application WO01/067103. Cells capable of responding to factors, simulating a state of interest are employed. Preferably the cells are primary cells in biologically relevant contexts. A sufficient number of factors are employed to involve a plurality of pathways and a sufficient number of parameters are selected to provide an informative dataset. The data resulting from the assays can be processed to provide robust comparisons between different environments and agents.
Among physiological states of interest are cardiovascular conditions. These conditions, including hypertension, atherosclerosis, and chronic heart failure affect more than 20% of the human population. Other conditions include angina, cardiac arrhythmia, cardiomyopathy, chronic venous insufficiency, diabetes, heart attack, high cholesterol, high homocysteine, high triglycerides, insulin resistance syndrome, and stroke. Inflammation has been associated with cardiovascular disease in several epidemiological studies, and the risk death from coronary disease has been shown to positively associate with elevated levels of inflammation markers, such as C-reactive protein (CRP), neopterin, matrix metalloproteinase-9 (MMP-9) and soluble intercellular adhesion molecules (sICAM).
Inflammatory cells, such as T cells, macrophages, etc., and inflammatory pathways, such as the NF-κB pathway, therefore contribute to the pathology of cardiovascular conditions. Inflammatory leukocyte trafficking into target tissues, e.g. atherosclerotic plaques, etc., and inflammatory mediators, e.g. cytokines, isoprenoids, etc. can affect cardiovascular processes including blood flow, vasoconstriction/dilation, and metabolic energy production.
However, while the cardiovascular stain drugs have been shown to have some immunomodulatory activities (see McCarey et al. (2004) Lancet 2004 363:2015-21), strategies to employ anti-inflammatory therapeutics for cardiovascular conditions have often met with failure (see Anker and Coats (2002) Int J Cardiol. 86:123-30; and Couzin (2004) Science 306:384-5).
Given the large number of cells and pathways involved in the development of cardiovascular inflammation, the evaluation of therapeutic modalities requires a complex assessment of effectiveness in multiple systems. The present invention addresses these issues.